[001] The present disclosure relates to the field of biopharmaceuticals and particularly refers to a composition exhibiting antibiofilm activity against the species of Candida genus. The present disclosure also discloses a process for preparing the composition.
BACKGROUND OF THE INVENTION
[002] Human skin naturally contains small amounts of microorganisms such as bacteria and fungi that are essential and are not harmful. But few bacteria and fungi can cause infections when begin to grow uncontrollably. One such potentially harmful fungi is the Candida species. Candida species are commensals in human microbiota that colonize in healthy individuals. But these species become opportunistic pathogens after the immunity of humans is compromised that leads to systemic-specific and invasive candidiasis. Candida infections are attributed to a high mortality rate of 46 -75 %, especially in invasive candidiasis reflecting the depth of severity in causing infection in both immunocompetent and immunocompromised patients. The most recurrently arising Candida infection in Intensive Care Unit (ICU) patients include mainly bloodstream, urinary tract, catheter-related and intra-abdominal infections. [003] The prevalent clinical related infections are primarily due to the members of the dimorphic Candida species such as C albicans andC tropicalis that can exist in yeast pseudohyphal or hyphal form. It has already been reported that yeast form of Candida species plays a vital role in the early stage of Candida infection (Saville et al., 2003 Engineered control of cell morphology in vivo reveals distinct roles for yeast and filamentous forms of Candida albicans during infection. Eukaryotic Cell. 2:1053-1060). The virulence factors such as biofilm formation, exoenzymatic factors such as phospholipase production, proteinase activity, coagulase activity and haemolysin production are crucial for Candida infection in human (Deorukhkar et al., 2014 Factors contributing to pathogenicity of Candida tropicalis and its antifungal susceptibility profile. International Journal of Microbiology. 13:144-152).

[004] There are wide varieties of broad-spectrum antifungal drugs such as azole drugs (Fluconazole, Itraconazole, Clotrimazole, and Ketoconazole), echinocandins (Micafungin, Caspofungin) and polyenes (Amphotericin B) which target specific virulence mechanisms of Candidiasis. One strategy of using antimicrobial has been disclosed in the patent document WO2018042367A2. The document discloses synthetic antimicrobial polymer-based compounds for managing microbial infections. In another patent document EP2365969B1, the novel agents exhibiting anti-biofilm activity have been disclosed. Particularly, the anti-biofilm agents thiazolidinedione and pyrrolidinedione derivatives are disclosed in the aforementioned document. Although, the antimicrobial drugs and the anti-biofilm agents shows effective anti-biofilm activity, however, the overuse of the aforementioned agents is accelerating the process of multi-drug resistance (Peman et al., 2009 Antifungal drug resistance mechanisms. Expert Review of Anti-infective Therapy. 7:453-460)
[005] Accordingly, there is a need for implementing the natural, non-toxic products that shows desired anti-biofilm activity and eliminate the emergence of multidrug resistance.
SUMMARY OF THE INVENTION
[006] In an aspect of the present disclosure, there is provided a composition comprising: a) tetradecanoic acid; and b) hexadecanoic acid, wherein tetradecanoic acid to hexadecenoic acid, wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006% with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01% with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1. [007] In another aspect of the present disclosure, there is provided a process for preparing the composition comprising: a) tetradecanoic acid; and b) hexadecanoic acid, wherein tetradecanoic acid to hexadecenoic acid, wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006%) with respect to the composition,

hexadecanoic acid is having a weight percentage in a range of 0.002-0.01% with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1, said process comprising: contacting tetradecanoic acid with hexadecanoic acid in presence of at least one solvent, at a speed in a range of 800-1200 rpm at a temperature in a range of 32°C to 40°C for a time period in a range of 5 to 20 minutes to obtain the composition.
[008] These and other features, aspects, and advantages of the present subject matter will be better understood with reference to the following description and appended claims. This summary is provided to introduce a selection of concepts in a simplified form. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used to limit the scope of the claimed subject matter.
BRIEF DESCRIPTION OF DRAWINGS
[009] The following drawings form a part of the present specification and are included to further illustrate aspects of the present disclosure. The disclosure may be better understood by reference to the drawings in combination with the detailed description of the specific embodiments presented herein
[010] Figure 1 depicts the percentage inhibition of biofilm formed by reference and clinically isolated Candida albicans in the presence of tetradecanoic acid (TA) at varying concentrations, in accordance with an embodiment of the present disclosure. [011] Figure 2 depicts the percentage inhibition of biofilm formed by reference and clinically isolated Candida tropicalis in the presence of tetradecanoic acid (TA) at varying concentrations, in accordance with an embodiment of the present disclosure. [012] Figure 3 depicts the percentage inhibition of biofilm formed by reference and clinically isolated Candida albicans in the presence of hexadecanoic acid (HA) at varying concentrations, in accordance with an embodiment of the present disclosure.

[013] Figure 4 depicts the percentage inhibition of biofilm formed by reference and clinically isolated Candida tropicalis in the presence of hexadecanoic acid (HA) at varying concentrations, in accordance with an embodiment of the present disclosure. [014] Figure 5 depicts the combinations of tetradecanoic acid (TA) & hexadecanoic acid (HA) used at different concentrations, in accordance with an embodiment of the present disclosure
DETAILED DESCRIPTION OF THE INVENTION
[015] Those skilled in the art will be aware that the present disclosure is subject to
variations and modifications other than those specifically described. It is to be
understood that the present disclosure includes all such variations and modifications.
The disclosure also includes all such steps, features, compositions, and compounds
referred to or indicated in this specification, individually or collectively, and any and
all combinations of any or more of such steps or features.
Definitions
[016] For convenience, before further description of the present disclosure, certain
terms employed in the specification, and examples are delineated here. These
definitions should be read in the light of the remainder of the disclosure and understood
as by a person of skill in the art. The terms used herein have the meanings recognized
and known to those of skill in the art, however, for convenience and completeness,
particular terms and their meanings are set forth below.
[017] The articles "a", "an" and "the" are used to refer to one or to more than one (i.e.,
to at least one) of the grammatical object of the article.
[018] The terms "comprise" and "comprising" are used in the inclusive, open sense,
meaning that additional elements may be included. It is not intended to be construed as
"consists of only".
[019] Throughout this specification, unless the context requires otherwise the word
"comprise", and variations such as "comprises" and "comprising", will be understood
5

to imply the inclusion of a stated element or step or group of element or steps but not
the exclusion of any other element or step or group of element or steps.
[020] The term "including" is used to mean "including but not limited to". "Including"
and "including but not limited to" are used interchangeably.
[021] The term "phytocompounds" (from Greek phyto, meaning "plant") are
chemicals produced by plants through primary or secondary metabolism. They
generally have biological activity in the plant host and play a role in plant growth or
defense against competitors, pathogens, or predators. The term "phytocompounds" can
be used interchangeably with "phytochemicals"
[022] The term "biofilm" is used to describe the aggregation of microorganisms in
which cells adhere to each other to a surface (living/nonliving). Fungal biofilms can be
formed on both abiotic and biotic surfaces. The biofilm formation confers numerous
advantages to the pathogens, including an increase in drug resistance and providing
high tolerance to environmental stresses such as antimicrobials and host immune
responses.
[023] The Biofilm Inhibitory Concentration (BIC) is defined as the lower
concentration of antibiotics/ antimicrobials showing 50% and 90% reduction of biofilm
binding. The percentage of biofilm inhibition is calculated by the following formula:
n, r.- «■■! i i -i -t- rAbsorbance of Control-Absorbance of Treated-, ,, __
% Biofilm Inhibition = f 1 x 100
k Absorbance of Control J
[024] As used herein, the term water-phase component and oil-phase component are collectively present as water in oil base components that are added to the active ingredients, forming the composition as disclosed in the present disclosure. Particularly, the water phase components are defined as components that dissolve in water. Similarly, oil phase components are defined as components that dissolve in oil. [025] For the purposes of the present document, the term "synergism" is used to describe the interaction or cooperation of two or more organizations, substances, or other agents to produce a combined effect greater than the sum of their separate effects. The synergistic effect shown by the combination of two phytochemicals at various

concentrations is studied by checker board assay through Fractional Inhibitory Concentration Index (FIC). Therefore, FIC is used to estimate the interaction between two or more phytochemicals used in combination.
i) If the FIC index is < 0.5, then the combination effect shown by the two or more
phytochemicals used in combination is denoted as synergistic.
ii) If the FIC index is between 0.5 to 2, then no interaction is shown by two or more phytochemicals used in combination.
iii) If the FIC index is > 2, then the combination effect shown by the two or more phytochemicals used in combination is denoted as antagonistic. [026] Ratios, concentrations, amounts, and other numerical data may be presented herein in a range format. It is to be understood that such range format is used merely for convenience and brevity and should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited.
[027] A wide variety of broad spectrum antifungal agents have been used to target specific virulence mechanisms of candidiasis. However, these antifungal agents show various adverse side effects and toxicity towards human cells at higher concentration. The side effects produced by the currently available antifungal drugs are associated with gastrointestinal, hepatic, and endocrinological disorders and interfere with oxidative drug metabolism in the liver of the host cells. In addition, the increasing incidence of drug-resistant pathogens and the toxicity of existing antifungal compounds has become an inevitable problem. In addition, the clinically isolated Candida species develops resistance against conventional antifungal agents due to the formation of biofilm structures. The ability of Candida to form biofilm causes various clinical problems of concern because they increase resistance to the antifungal therapies. The rising concern on side effects of antifungals and multidrug resistant pathogen is curated by plant based active principles. Therefore, the increasing incidence of drug-resistant

pathogens and the toxicity of existing antifungal compounds have drawn attention towards the antimicrobial activity of natural products.
[028] To overcome the problem of side effects produced by the antifungal agents, and to reduce the probability of developing multidrug resistance by the pathogens against the antifungal agents, the present disclosure deploys a combinatorial strategy of developing an effective, non- toxic, combination of natural components against the biofilm formed by Candida infection.
[029] The present disclosure relates to the combinatorial strategy of using plant based phytocompounds (phytochemicals). Particularly, the present disclosure discloses a composition comprising phytochemicals such as tetradecanoic acid (TA) and hexadecanoic acid (HA), wherein tetradecanoic acid has a weight percentage in a range of 0.001-0.006% with respect to the composition, hexadecanoic acid has a weight percentage in a range of 0.002-0.01% with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1. It is specified that the composition provides synergistic inhibitory activity against biofilm formation in dimorphic species of Candida genus such as Candida albicans and Candida tropicalis. Additionally, the synergistic combination of tetradecanoic acid (TA) and hexadecanoic acid (HA) exhibits antibiofilm efficacy at lower concentrations without exerting toxicity to the growth of Candida species. Further, this combinatorial strategy helps in decreasing the probability of developing drug resistance. [030] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the disclosure, the preferred methods, and materials are now described. All publications mentioned herein are incorporated herein by reference.
[031] The present disclosure is not to be limited in scope by the specific embodiments described herein, which are intended for the purposes of exemplification only.

Functionally-equivalent products, compositions, and methods are clearly within the scope of the disclosure, as described herein.
[032] In an embodiment of the present disclosure, there is provided a composition comprising: a) tetradecanoic acid; and b) hexadecanoic acid, wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006% with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01% with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1. In another embodiment of the present disclosure, tetradecanoic acid to hexadecanoic acid has a weight ratio in the range of 0.620:1 to 1:1. In yet another embodiment of the present disclosure, tetradecanoic acid to hexadecanoic acid has a weight ratio in the range of 1:1 to 1:1. [033] In an embodiment of the present disclosure, there is provided a composition comprising: a) tetradecanoic acid; and b) hexadecanoic acid, wherein tetradecanoic acid is having a weight percentage in a range of 0.002-0.006%) with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.009%) with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1.
[034] In an embodiment of the present disclosure, there is provided a composition comprising: a) tetradecanoic acid , b) hexadecanoic acid and c) at least one water phase component, wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006%) with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01%) with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1. [035] In an embodiment of the present disclosure, there is provided a composition comprising: a) tetradecanoic acid, b) hexadecanoic acid, and c) at least one oil phase component, wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006%) with respect to the composition, hexadecanoic acid is having a weight

percentage in a range of 0.002-0.01% with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1. [036] In an embodiment of the present disclosure, there is provided a composition comprising: a) tetradecanoic acid, b) hexadecanoic acid, and c) a combination of at least one water phase and at least one oil phase component, wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006% with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01%) with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1.
[037] In an embodiment of the present disclosure, there is provided a composition comprising: a) tetradecanoic acid, b) hexadecanoic acid, and c) at least one water phase component , wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006%) with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01%) with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1 and wherein the at least one water phase component is selected from a group consisting of hydrosols, humectants, and combinations thereof.
[038] In an embodiment of the present disclosure, there is provided a composition comprising: a) tetradecanoic acid, b) hexadecanoic acid and c) at least one oil phase component , wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006%) with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01%> with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1 and wherein the at least one oil phase component is selected from a group consisting of preservatives, emollients, structurants, thickeners, emulsifiers, and combinations thereof.
[039] In an embodiment of the present disclosure, there is provided a composition comprising: a) tetradecanoic acid, b) hexadecanoic acid and c) a combination of at least

one water phase and at least one oil phase component, wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006% with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01% with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1 , and wherein the at least one water phase component is selected from a group consisting of hydrosols, humectants, and combinations thereof and wherein the at least one oil phase component is selected from a group consisting of preservatives, emollients, structurants, thickeners, emulsifiers, and combinations thereof.
[040] In an embodiment of the present disclosure, there is provided a composition comprising: a) tetradecanoic acid b) hexadecanoic acid, c) at least one water phase component , wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006%) with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01%) with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1, wherein the at least one water phase component is selected from a group consisting of hydrosols, humectants, and combinations thereof and wherein the hydrosols has a weight percentage in a range of 50-80%> with respect to the composition, humectants has a weight percentage in a range of 1-10% with respect to the composition. [041] In an embodiment of the present disclosure, there is provided a composition comprising: a) tetradecanoic acid b) hexadecanoic acid and c) at least one oil phase component, wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006%) with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01%) with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1, and wherein the at least one oil phase component is selected from a group consisting of preservatives, emollients, structurants, thickeners, emulsifiers, and combinations thereof and wherein the preservatives has a weight percentage in range of 0.001-0.5%)

with respect to the composition, emollients has a weight percentage in a range of 0.05-20% with respect to the composition, structurants has a weight percentage in a range of 0.01-7% with respect to the composition, thickeners has a weight percentage in a range of 0.05 to 5%> with respect to the composition and emulsifiers has a weight percentage in a range of 1 to 10% with respect to the composition.
[042] In an embodiment of the present disclosure, there is provided a composition comprising: a) tetradecanoic acid b) hexadecanoic acid and c) a combination of at least one water phase and at least one oil phase component, wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006%) with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01%) with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1, and wherein the at least one water phase component is selected from a group consisting of hydrosols, humectants, and combinations thereof, and wherein the hydrosols has a weight percentage in a range of 50-80%) with respect to the composition, humectants has a weight percentage in a range of 1-10%> with respect to the composition, and wherein the at least one oil phase component is selected from a group consisting of preservatives, emollients, structurants, thickeners, emulsifiers, and combinations thereof and wherein the preservatives has a weight percentage in range of 0.001-0.5%) with respect to the composition, emollients has a weight percentage in a range of 0.05-20%) with respect to the composition, emollients has a weight percentage in a range of 0.05-20%> with respect to the composition, structurants has a weight percentage in a range of 0.01-7% with respect to the composition, thickeners has a weight percentage in a range of 0.05 to 5% with respect to the composition and emulsifiers has a weight percentage in a range of 1 to 10%> with respect to the composition.
[043] In an embodiment of the present disclosure, there is provided a composition comprising: a) tetradecanoic acid b) hexadecanoic acid and c) at least one water phase, wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006%)

with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01% with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1, and wherein the at least one water phase component is selected from a group consisting of hydrosols, humectants, and combinations thereof. In another embodiment of the present disclosure, the hydrosols are selected from a group consisting of apple, artemisia, catnip, chamomile, clari sage, ginger root, herbal tulsi basil, lavender, lemongrass, mount mint, pachouli, rosewood, sandalwood distillate, spearmint and combinations thereof , having a weight percentage in the range of 50-80% with respect to the composition and the humectants are selected from a group consisting of aloe vera powder, arnica extract, algae extract, hydrolyzed baobab protein, coenzyme qlO, hydrolyzed collagen protein, colloidal oatmeal, glycerin, caprylyl glycol, ceramide complex, lacto-ceramide, matrixyl s-6, propylene glycol, rhubarb root extract, hyaluronic acid, hydrolyzed jojoba protein, hydrolyzed keratin protein, sodium PC A and combinations thereof, having a weight percentage in the range of 1 to 10% with respect to the composition.
[044] In an embodiment of the present disclosure, there is provided a composition comprising: a) tetradecanoic acid b) hexadecanoic acid and c) at least one oil phase component, wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006%) with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01%) with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1, and wherein the at least one oil phase component is selected from a group consisting of preservatives, emollients, structurants, thickeners, emulsifiers, and combinations thereof. In another embodiment of the present disclosure, the preservatives are selected from a group consisting of paraben, butylatedhydroxyanisole, gluconolactone, sodium benzoate, chlorobutanol, benzyl alcohol, sodium sorbate, imidazolidinyl urea, benzalkonium chloride, alpha tocopherol, methyl-dibromo glutaronitrile, tea tree

essential oil, having a weight percentage in range of 0.001-0.5% with respect to the composition, the emollients are selected from a group consisting of amodimethicone, carnaubawax, cetyl alcohol, cetyl ester wax, emulsifying wax, hydrous lanolin, lanolin, lanolin alcohols, microcrystalline wax, paraffin, octyldodecanol, amodimethicone, cyclodimethicone, cyclomethicone, dimethicone 500, dimethicone satin, isodimethicone copolymer, PEG8, polyisobutene 250, squalane, almond oil, castor oil, grapeseed oil, red raspberry seed oil, cherry kernel oil having a weight percentage in a range of 0.05-20% with respect to the composition, the structurants are selected from a group consisting of rice bran oil, corn oil, sunflower oil, starch, alginate, bees wax, berry wax, fruit wax, silicone based polymers, acrylate copolymer, cellulose based polymers, having a weight percentage in a range of 0.01-7%> with respect to the composition, the thickeners are selected from a group consisting of carbomer, methyl cellulose, sodium carboxyl methyl cellulose, carrageenan, colloidal silicon dioxide, guar gum, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, gelatin, polyethylene oxide, alginic acid, sodium alginate, fumed silica having a weight percentage in a range of 0.05 to 5%> with respect to the composition and the emulsifiers are selected from a group consisting of polysorbate 20, polysorbate 80, polysorbate 60, poloxamer, emulsifying wax, sorbitan monostearate, sorbitan monooleate, sodium lauryl sulfate, propylene glycol monostearate, diethylene glycol monoethyl ether, docusate sodium having a weight percentage in a range of 1 to 10%> with respect to the composition.
[045] In an embodiment of the present disclosure, there is provided a process for preparing the composition comprising: a) tetradecanoic acid; and b) hexadecanoic acid, wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006%) with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01%) with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1, said process comprising contacting tetradecanoic acid with hexadecanoic acid in presence of at least

one solvent, at a speed in a range of 800-1200 rpm at a temperature in a range of 32°C to 40°C for a time period in a range of 5 to 20 minutes to obtain the composition. [046] In an embodiment of the present disclosure, there is provided a process for preparing the composition: a) tetradecanoic acid; b) hexadecanoic acid; and (c) a combination of at least one water phase and at least one oil phase component, said process comprising: (i) contacting tetradecanoic acid with hexadecanoic acid in presence of an oil phase component at a speed in a range of 400-800 rpm for a time in range of 10-15 minutes at a temperature in a range of 35-40°C to obtain a first mixture; ii) contacting at least one water phase component with at least one oil phase component at a speed in a range of 900-1100 rpm for a time period in a range of 5-10 minutes at a temperature in a range of 40-45°C, to obtain a second mixture; and iii) contacting the first mixture and the second mixture at a speed in a range of 900-1100 rpm for a time period in a range of 2-10 minutes at a temperature in a range of 40-45°C to obtain the composition.
[047] In an embodiment of the present disclosure, there is provided a process for preparing the composition comprising: a) tetradecanoic acid; and b) hexadecanoic acid, wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006% with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01% with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1, said process comprising contacting tetradecanoic acid with hexadecanoic acid in presence of at least one solvent, at a speed in a range of 800-1200 rpm at a temperature in a range of 32°C to 40°C for a time period in a range of 5 to 20 minutes to obtain the composition, and wherein the at least one solvent is selected from a group consisting of water, dimethyl sulphoxide (DMSO), isopropanol, chloroform, hexane, carbon disulphide and combinations thereof. In another embodiment of the present disclosure, the at least one solvent is water. In yet another embodiment of the present disclosure, the at least one solvent is dimethyl sulphoxide.

[048] In an embodiment of the present disclosure, there is provided a process for preparing the composition: a) tetradecanoic acid; b) hexadecanoic acid; and (c) a combination of at least one water phase and at least one oil phase component, said process comprising: (i) contacting tetradecanoic acid with hexadecanoic acid in presence of an oil phase component at a speed in a range of 400-800 rpm for a time in range of 10-15 minutes at a temperature in a range of 35-40°C to obtain a first mixture; ii) contacting at least one water phase component with at least one oil phase component at a speed in a range of 900-1100 rpm for a time period in a range of 5-10 minutes at a temperature in a range of 40-45°C, to obtain a second mixture; and iii) contacting the first mixture and the second mixture at a speed in a range of 900-1100 rpm for a time period in a range of 2-10 minutes at a temperature in a range of 40-45°C to obtain the composition, and wherein the at least one solvent is selected from a group consisting of water, dimethyl sulphoxide (DMSO), isopropanol, chloroform, hexane, carbon disulphide and combinations thereof.
[049] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein said composition exhibits inhibitory activity against biofilm formation in species of Candida genus.
[050] In an embodiment of the present disclosure, there is provided a formulation comprising (a) tetradecanoic acid; (b) hexadecanoic acid; and (c) excipient, wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006% with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01% with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1, and wherein the excipient is selected from the group consisting of at least one water phase component, at least one oil phase component, and combinations thereof, and wherein the at least one water phase component selected from a group consisting of hydrosols having a weight percentage in a range of 50-80%) with respect to the composition, humectants having a weight percentage in a range of 1-10% with respect to the composition, and combinations

thereof and the at least one oil phase component is selected from a group consisting of preservatives having a weight percentage in range of 0.001-0.5% with respect to the composition, emollients having a weight percentage in a range of 0.05-20% with respect to the composition,, structurants having a weight percentage in a range of 0.01-7%> with respect to the composition, thickeners having a weight percentage in a range of 0.05 to 5%>, emulsifiers having a weight percentage in a range of 1 to 10% with respect to the composition, and combinations thereof. In another embodiment of the present disclosure, the formulation is in a form of ointment. In yet another embodiment of the present disclosure, the formulation is in a form of lotion. [051] Although the subject matter has been described in considerable detail with reference to certain examples and implementations thereof, other implementations are possible.
EXAMPLES
[052] The disclosure will now be illustrated with working examples, which is intended to illustrate the working of disclosure and not intended to take restrictively to imply any limitations on the scope of the present disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods, devices and materials are described herein. It is to be understood that this disclosure is not limited to particular methods, and experimental conditions described, as such methods and conditions may apply.
[053] The working examples as depicted in the forthcoming sections highlight the combinatorial effect of different phytochemicals used at different concentrations on C. albicans and C. tropicalis in achieving a composition. The composition as disclosure in the present disclosure comprising: a) tetradecanoic acid; and b) hexadecanoic acid,

wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006% with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01% with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1 exhibits inhibitory activity against biofilm formation in species of Candida genus such as C albicans and C tropicalis. It is further specified that the presence of tetradecanoic acid and hexadecenoic acid in their disclosed ranges is critical to achieve synergistic effect on inhibiting the biofilm formed by C. albicans and C. tropicalis. Also, the antibiofilm efficacy exhibited by the composition even at low concentration shows that the composition is non-toxic to the species of Candida genus and further reduces the probability of developing multidrug resistance. When any of the components as specified above are present outside the disclosed ranges then it would substantially affect the synergistic anti-biofilm activity against the species of species of Candida genus.
Materials and Methods
[054] The phytochemicals such as tetradecanoic acid (TA) and hexadecanoic acid (HA) were used for arriving at the composition of the present disclosure. Tetradecanoic acid and hexadecanoic acid were procured from Sigma-Aldrich and TCI limited, respectively. The antibiofilm efficacy of the phytochemicals as mentioned above were studied against the species of Candida genus forming biofilm structures. Candida strains used in this study are as given below:

|055J All strains were maintained in Sabouraud Dextrose Agar medium (HI Media Labs). The cells were grown in YEPD medium (yeast extract-1%, peptone-2% and dextrose-2%, purchased from HI Media Labs) for 12 h at 37°C with agitation and 1% (V/V) of 12 h grown culture was used for biofilm studies. In biofilm assay, the cells were grown in spider medium (mannitol-1%, K2HPO4 - 0.25% and nutrient broth -1%, purchased from Hi Media Labs)
Example 1
Evaluation of Biofilm Inhibitory Concentration (BIC) of TA and HA against biofilm formed by C. albicans and C. tropicalis
[056] Different concentrations of the components (phytochemicals) such as TA and HA were used against the species of Candida genus. The effect of components TA and HA on biofilm formed by C. albicans and C. tropicalis at different concentrations were evaluated to determine the Biofilm Inhibitory Concentration (BIC) of the respective components. The BIC of TA and HA against C. albicans and C. tropicalis biofilm was evaluated by the following steps: C. albicans and C. tropicalis were allowed to form biofilm on 24 well Micro Titer Plate (MTP) in spider medium without and with TA at concentrations: 15.625 31.25, 62.5, 125 and 250 ugmL"1 and HA at concentrations: 25, 50, 100 and 200 ug mL"1. The 24 well Micro Titer Plate (MTP) was incubated at 37° C for 24 h. After incubation, the plate was washed with distilled water to remove unbound cells. Subsequently, the bound cells in the plate were stained with crystal violet followed by destaining with 10% glacial acetic acid. The difference in biofilm formation was measured spectrophotometrically at 570 nm. The percentage of biofilm inhibition was calculated by the following formula:

[057] Checkerboard assay - Checkerboard assay is an approach of studying the effect of the potency of the combination of phytochemicals in comparison to the effect of potency of individual phytochemical. The effect of the potency of the combination of phytochemicals in comparison to the effect of potency of individual phytochemical is determined by fractional inhibitory concentration (FIC) index. The phytochemical combinatorial effect is defined by the FIC index value.
• If FIC index is <0.5, then the phytochemical combinatorial effect is denoted as synergistic effect.
• If FIC index is between 0.5 to 2.0, then the phytochemical combinatorial effect is denoted as indifferent.
• If FIC index is >2.0, then the phytochemical combinatorial effect is denoted as antagonistic effect
[058] In the present disclosure, the combinatorial effect of tetradecanoic acid and hexadecanoic acid was evaluated by using Checkerboard assay. Tetradecanoic acid and hexadecanoic acid were used in a combination to inhibit the biofilm formed by the species of Candida genus. To study the effectiveness of the combination of the aforementioned components, FIC index value was determined to describe the combinatorial effect of TA and HA against biofilm structures formed by C. albicans and C. tropicalis. The FIC index was determined as prescribed by Odds, 2003 as follows:

[059] The effect of TA on C. albicans and C. tropicalis was studied using varying concentrations of TA individually. Particularly, by using the various concentrations of TA, the biofilm structure formed by both reference and clinically isolated C. albicans and C. tropicalis were inhibited. Further, the biofilm structure formed by C. albicans and C. tropicalis were inhibited in a concentration dependent manner without inhibiting the growth of the species of Candida genus. The effect of TA on C. albicans and C. tropicalis was studied by using TA at different concentrations such as 0, 15.625, 31.25, 62.5, 125 and 250 ug ml/1.
[060] Figure 1 illustrates the percentage inhibition of biofilm formed by both reference and clinically isolated C. albicans in the presence of TA at different concentrations. Referring to the Figure 1, maximum inhibition of C. albicans biofilm was observed by using TA at a concentration of 125 ug mL"1. Figure 2 illustrates the percentage inhibition of biofilm formed by both reference and clinically isolated C. tropicalis in the presence of TA at different concentrations. Referring to the Figure 2, maximum inhibition of reference strain, CI 1 and CI 3 biofilm was observed at a concentration of 125 ug mL_1and in case of CI 2, the inhibition was observed at a concentration of 250 ug mL"1 of TA.
(b) Effect of HA on C. albicans and C. tropicalis [061] The effect of HA on C. albicans and C. tropicalis was studied using varying concentrations of HA individually. Particularly, by using the various concentrations of HA, the biofilm structure formed by both reference and clinically isolated C. albicans

and C tropicalis were inhibited. Further, the biofilm structure formed by C albicans and C. tropicalis were inhibited in a concentration dependent manner without inhibiting the growth of the species of Candida genus. The effect of HA on C. albicans and C. tropicalis was studied by using TA at different concentrations such as 25, 50, 100, 200 and 400 ugml/1.
[062] Figure 3 illustrates the percentage inhibition of biofilm formed by both reference and clinically isolated C. albicans strains in the presence of HA at different concentrations. Referring to the Figure 3, the maximum inhibition of biofilm formed by reference strains such as CI 4 and CI 6 was observed by using HA at a concentration of 200 ug ml/1 and in clinically isolated strains such as CI 5, the maximum inhibition of biofilm was observed at a concentration of 400 ug ml/1 of HA. [063] Figure 4 illustrates the percentage inhibition of biofilm formed by both reference and clinically isolated C tropicalis strains in the presence of HA at different concentrations. Referring to the Figure 4, maximum inhibition of biofilm formed by reference strains such as CI 1 and CI 3 was observed at a concentration of 200 ug ml/ 1 and in case of CI 2, the maximum inhibition of biofilm was observed at a concentration of 400 ug ml/1 of TA.
Example 2
Effect of composition of the present disclosure on biofilm formed by the reference
stains and clinically isolated strains of C. albicans and C. tropicalis
[064] Various combinations of TA and HA were used against biofilm produced by
different strains of C albicans and C tropicalis. The combinatorial effect of TA and
HA against biofilm produced by C albicans and C tropicalis was evaluated by
checkerboard assay (as explained in Example 1 above). Figure 5 illustrates the various
combinations of TA and HA at different concentrations utilized in this study. This
layout of combination was followed for both C albicans and C tropicalis biofilm
inhibition study.

a) Evaluation of the composition on biofilm formed by C. albicans (ATCC): [065] Combinations of components such as TA and HA at different concentrations were utilized to assess their antibiofilm activity against ATCC strain of C. albicans, and their activity was evaluated by using checkerboard assay. Table 1 depicts the fractional inhibitory concentration (FIC) indices of combinations of TA and HA at different concentrations for assessing their antibiofilm activity. As depicted in the Table 1, the signs ' + ' and ' - ' indicates the presence and absence of antibiofilm activity, respectively in individual and combination of components. The sign ' * ' indicates the synergistic combination of TA+HA showing antibiofilm activity against ATCC strain of C. albicans.
[066] Referring to Table 1, it was observed that TA and HA when used at a combination of 31.25 + 25 ug mL"1, 31.25 + 50 ug mL"1 showed synergistic antibiofilm activity against ATCC strain of C. albicans, with FIC indices of 0.375, and 0.5, respectively. Although, the combination of TA and HA at concentration of 15.625 ug
23

0.375, respectively, however, the combination at the atorementioned concentrations did not show the desired antibiofilm activity against ATCC strain of C. albicans. [067] It was also observed that none of the combinations of TA + HA with TA having concentrations of 62.5, 125, and 250 ug mL"1 and HA having concentrations of 100, 200, 400 ug mL"1 showed synergistic antibiofilm activity against ATCC strain of C. albicans, with their FIC indices being greater than 0.5.
[068] It can also be inferred from the above observations that HA and TA when present in the disclosed weight ranges of the present disclosure to form combinations of 31.25 + 25 ug mL"1 and 31.25 + 50 ug mL"1, showed desired synergistic antibiofilm activity against ATCC strain of C. albicans, whereas when TA and HA were present outside the disclosed weight ranges at concentrations such as 62.5,125, 250 ug mL"1 and 100, 200, 400 ug mL"1, respectively, showed no antibiofilm activity against ATCC strain of C. albicans,.
b) Evaluation of composition on biofilm formed by clinically isolated C. albicans (CI 4):
[069] Combinations of components such as TA and HA at different concentrations were utilized to assess their antibiofilm activity against clinically isolated CI 4 strain of C. albicans. Table 2 depicts the Fractional Inhibitory Concentration (FIC) indices of TA and HA combinations evaluated at different concentrations by using checkerboard assay to check their antibiofilm activity against clinically isolated clinically isolated CI 4 strain of C. albicans. As depicted in the Table 2, the signs ' + ' and ' -' indicate the presence and absence of antibiofilm activity, respectively in individual and combination of components. The sign ' * ' indicates the synergistic combination of TA+HA showing antibiofilm activity against clinically isolated CI 4 strain of C. albicans. Table 2

[070] It can be observed from Table 2 that the TA and HA when used at a combination of 31.25 + 50 ug mL"1 showed the synergistic antibiofilm activity against CI 4 strain of C. albicans, with FIC index of 0.5. It was further observed that the combination of TA + HA used at various concentrations such as 62.5, 125, 250 ug mL"1 of TA and 100, 200, 400 ug mL"1 of HA did not show the synergistic effect against CI 4 strain of C. albicans, with FIC indices greater than 0.5
(c) Evaluation of the composition in inhibiting the biofilm formation by
clinically isolated C. albicans (CI 5) biofilm [071] Combinations of components such as TA and HA at different concentrations were utilized to assess their antibiofilm activity against CI 5 strain of C. albicans, and their activity was evaluated by using checkerboard assay. Table 3 depicts the Fractional Inhibitory Concentration (FIC) indices of TA and HA combinations at different concentrations to evaluate their synergistic antibiofilm activity against clinically isolated CI 5 strain of C. albicans. As depicted in the Table 3, the signs ' + ' and ' -'

indicate the presence and absence of antibiofilm activity, respectively in individual and combination of components. The sign ' * ' indicates the synergistic antibiofilm activity exhibited by the combination of TA+HA.
[072] It was observed that the TA and HA when used at a combination of 15.625+100, 31.25 + 25, 31.25 + 50 and 31.25 + 100 ug ml/1 showed the synergistic effect in inhibiting the biofilm formed by CI 5 strain of C. albicans, with FIC indices of 0.375, 0.3125, 0.375 and 0.5, respectively.
[073] Further, it was observed that the combination of TA + HA used at various concentrations such as 62.5, 125, 250 ug ml/1 of TA and 200, 400 ug mL"1 of HA did not show the synergistic effect in inhibiting the biofilm formed by CI 5 strain of C. albicans, with FIC indices greater than 0.5.
(d) Evaluation of composition on biofilm formed by clinically isolated C. albicans (CI 6):

[074] Combinations of components such as TA and HA at different concentrations were utilized to assess their antibiofilm activity against clinically isolated CI 6 strain of C. albicans. Table 4 depicts the Fractional Inhibitory Concentration (FIC) indices of TA and HA combinations at different concentrations to check their antibiofilm activity against clinically isolated CI 6 strain of C. albicans. As depicted in the Table 4, the signs ' +' and ' -' indicate the presence and absence of antibiofilm activity, respectively in individual and combination of components. The sign ' * ' indicates the synergistic combination of TA+HA showing antibiofilm activity against clinically isolated CI 6 strain of C. albicans.
[075] It was observed from Table 4, that the TA and HA when used at a combination of 15.625 + 50, 31.25 + 25 and 31.25 + 50 ug ml/1 showed synergistic antibiofilm activity against CI 6 strain of C. albicans, with FIC indices of 0.375, 0.375 and 0.5, respectively.

[076] Further, it was observed that the combination of TA + HA used at various concentrations such as 62.5, 125, 250 ug ml/1 of TA and 100, 200, 400 ug ml/1 of HA did not show the synergistic antibiofilm activity against CI 6 strain of C albicans, with FIC indices greater than 0.5.
e) Evaluation of composition on biofilm formed by C.tropicalis (MTCC): [077] Combinations of components such as TA and HA at different concentrations were utilized to assess their antibiofilm activity against MTCC strain of C.tropicalis. Table 5 shows the Fractional Inhibitory Concentration (FIC) indices of TA and HA combinations evaluated at different concentrations by using checkerboard assay to check their antibiofilm activity against MTCC strain of C.tropicalis. As shown in the Table 5, the signs ' +' and' -' indicate the presence and absence of antibiofilm activity, respectively in individual and combination of components. The sign ' * ' indicates the synergistic combination of TA + HA showing antibiofilm activity against MTCC strain of C.tropicalis .

[078] Referring to Table 5, it can be observed that the TA and HA used at a combination of 15.625 + 50, 31.25 + 25 and 31.25 + 50 ug mL"1 showed synergistic antibiofilm activity against MTCC strain of C.tropicalis, with FIC indices of 0.375, 0.375 and 0.5, respectively.
[079] Further, it was observed that the combination of TA + HA used at various concentrations such as 62.5, 125, 250 ug mL"1 of TA and 100, 200, 400 ug mL"1 of HA did not show the synergistic effect in inhibiting the biofilm formed by MTCC strain of C.tropicalis, with FIC indices greater than 0.5.
f) Evaluation of composition on biofilm formed by clinically isolated C.tropicalis (CI 1) biofilm:
[080] Combinations of components such as TA and HA at different concentrations were utilized to assess their antibiofilm activity against CI 1 strain of C tropicalis. Table 6 depicts the Fractional Inhibitory Concentration (FIC) indices of TA and HA combinations evaluated at different concentrations by using checkerboard assay to check their antibiofilm activity against clinically isolated CI 1 strain of C.tropicalis. As depicted in the Table 6, the signs ' + ' and ' - ' indicate the presence and absence of antibiofilm activity, respectively in individual and combination of components. The sign ' * ' indicates the synergistic combination of TA+HA showing antibiofilm activity against CI 1 strain of C.tropicalis. Table 6

[081] Referring to Table 6, it can be observed that the TA and HA when used at a combination of 31.25+50 ug mL"1 showed synergistic effect in inhibiting the biofilm formed by CI 1 strain of C.tropicalis, with FIC index of 0.5.
[082] Further, it was observed that the combination of TA + HA used at various concentrations such as 62.5, 125, 250 ug mL"1 of TA and 100, 200, 400 ug mL"1 of HA did not show the synergistic effect in inhibiting the biofilm formed by CI 1 strain of C.tropicalis, with FIC indices greater than 0.5.
g) Evaluation of composition on biofilm formed by clinically isolated C.tropicalis (CI 2) biofilm:
[083] Combinations of components such as TA and HA at different concentrations were utilized to assess their antibiofilm activity against CI 2 strain of C tropicalis. Table 7 depicts the Fractional Inhibitory Concentration (FIC) indices of TA and HA combinations evaluated at different concentrations by using checkerboard assay to check their antibiofilm activity against clinically isolated CI 2 strain of C.tropicalis. As depicted in the Table 7, the signs ' + ' and ' - ' indicate the presence and absence of

[084] Referring to Table 7, it was observed that the TA and HA used at a combination of 15.625 + 100, 31.25 + 50 and 31.25 + 100 showed synergistic antibiofilm activity against CI 2 strain of C.tropicalis, with FIC indices of 0.375, 0.375 and 0.5, respectively.
[085] Further, it was observed that the combination of TA + HA used at various concentrations such as 62.5, 125, 250 ug ml/1 of TA and 200, 400 ug mL"1 of HA did not show the synergistic antibiofilm activity against CI 2 strain of C. tropicalis, with FIC indices greater than 0.5.

(h) Evaluation of composition on biofilm formed by clinically isolated C.tropicalis (CI 3):
[086] Combinations of components such as TA and HA at different concentrations were utilized to assess their antibiofilm activity against clinically isolated CI 3 strain of C.tropicalis. Table 8 shows the fractional inhibitory concentration (FIC) indices of TA and HA combinations evaluated at different concentrations by using checkerboard assay to check their antibiofilm activity against clinically isolated CI 3 strain of C.tropicalis. As depicted in the Table 8, the signs ' + ' and ' -' indicate the presence and absence of antibiofilm activity, respectively in individual and combination of components. The sign ' * ' indicates the synergistic combination of TA+HA showing antibiofilm activity against clinically isolated CI 3 strain of C tropicalis.
[087] Referring to Table 8, it can be observed that the TA and HA when used at a combination of 31.25 + 50 ug ml/1 showed the synergistic effect in inhibiting the antibiofilm formed by CI 3 strain of C.tropicalis, with FIC index of 0.5.

[088] Further, it was observed that the combination of TA + HA used at various concentrations such as 62.5, 125, 250 ug mL"1 of TA and 25, 50, 100, 200, 400 ug mL" 1 of HA did not show the synergistic effect in inhibiting the antibiofilm formed by CI 3 strain of Ctropicalis, with FIC indices greater than 0.5.
[089] Overall, it can be inferred from Table 1-8 that the combination of the phytochemicals such as tetradecanoic acid and hexadecenoic acid in the disclosed weight percentage range of 0.001-0.006% and 0.002-0.01%, respectively, and weight ratio range (0.350:1 to 1:1) is essential to arrive at the composition of the present disclosure so as to have a desired synergistic effect against the biofilm formation by the species of Candida genus such as C. albicans and C. tropicalis. The antibiofilm activity exhibited by TA and HA, individually and in combinations thereof against the species of Candida genus are summarised in Table 9 below.

[090] On comparing the biofilm inhibitory effect of the components TA and HA, individually and in combinations thereof as provided in Table 9, it was observed that TA at a 125 ug mL"1 and 250 ug mL"1 showed antibiofilm effect against biofilm formation by the species of C. albicans and C. tropicalis, respectively, and HA at a concentration of 200 and 400 ug mL"1 showed antibiofilm effect against biofilm formation by the species of C. albicans and C. tropicalis, respectively. However, TA and HA when used in combination showed synergistic antibiofilm activity against the species of C. albicans and C. tropicalis at low concentrations, i.e. 15.625+100, 31.25+25, 31.25+50 and 31.25+100 ug ml/1.
Example 3
Process for preparing the composition of the present disclosure
[091] The process for preparing the composition of the present disclosure comprised: Tetradecanoic acid was contacted with hexadecanoic acid in presence of purified water and Dimethyl Sulphoxide (DMSO) at stock concentration of 0.05 %, at a speed in a range of 800-1200 rpm at a temperature in a range of 32°C to 40°C for a time period in a range of 5 to 20 minutes to obtain the composition. Example 4

Process for preparing the composition having a combination of water phase and oil phase component of the present disclosure
[092] The composition of the present disclosure was utilized for preparing the formulation that can be used in the form of topical cream and topical lotion. The formulation comprising the composition further comprised water phase component and oil phase component.
[093] Water phase components utilised in the present disclosure are as
follows:
1. Any one of the hydrosols such as apple, artemisia, catnip, chamomile, clari sage, ginger root, herbal tulsi basil, lavender, lemongrass, mount mint, pachouli, rosewood, sandalwood distillate and spearmint acts as a skin care and can be used at the range of 50 - 80 %.
2. Any one of the humectants such as aloe vera powder, arnica extract, algae extract, hydrolyzed baobab protein, coenzyme qlO, hydrolyzed collagen protein, colloidal oatmeal, glycerin, caprylyl glycol, ceramide complex, lacto-ceramide, matrixyl s-6, propylene glycol, rhubarb root extract, hyaluronic acid, hydrolyzed jojoba protein, hydrolyzed keratin protein, sodium PC A can be used at the range of 1 % to 10% that promotes the retention of water in the lotion/ointment system.
[094] Oil phase components utilised in the present disclosure are as follows:
1. Any one of the preservatives such as paraben, butylatedhydroxyanisole,
gluconolactone, sodium benzoate, chlorobutanol, benzyl alcohol, sodium
sorbate, imidazolidinyl urea, benzalkonium chloride, alpha tocopherol, methyl-
dibromo glutaronitrile, tea tree essential oil prevents microbial contamination
in the lotion/ointment and can be used at the range of 0.001 % to 0.5%.
2. Any one of the emollients form such as amodimethicone, carnauba wax, cetyl
alcohol, cetyl ester wax, emulsifying wax, hydrous lanolin, lanolin, lanolin

alcohols, microcrystalline wax, paraffin,, octyldodecanol, amodimethicone, cyclodimethicone, cyclomethicone, dimethicone 500, dimethicone satin, isodimethicone copolymer, PEG8, polyisobutene 250, squalane, almond oil, castor oil, grapeseed oil, red raspberry seed oil, cherry kernel oil forms the basic structure for semisolid ointment and can be used at the range of 0.05% to 20%.
3. Any one of the structurants such as rice bran oil, corn oil, sunflower oil, starch, alginate, bees wax, berry wax, fruit wax, silicone-based polymers, acrylate copolymer, cellulose based polymers can be used at the range of 0.01 to 7%> to enhance the texture of lotion/ointment.
4. Any one of the thickeners such as carbomer, methyl cellulose, sodium carboxyl methyl cellulose, carrageenan, colloidal silicon dioxide, guar gum, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, gelatin, polyethylene oxide, alginic acid, sodium alginate, fumed silica can be used in the range of 0.05 to 5 % to increase the viscosity of base materials in lotion/ointment.
5. Any one of the emulsifiers such as polysorbate 20, polysorbate 80,
polysorbate 60, poloxamer, emulsifying wax, sorbitan monostearate, sorbitan
monooleate, sodium lauryl sulfate, propylene glycol monostearate, diethylene
glycol monoethyl ether, docusate sodium can be used in the range of 1 to 10 %
to enhance the solubility of hydrophobic particles.
[095] Overall the components used for preparing the formulation are shown in Table 10 below.

[096] The formulation of the synergistic combination of tetradecanoic acid and 3-hexadecanoic acid as disclosed in the present disclosure was prepared by the following steps:
1. The active ingredients in the current invention i.e. TA and HA were prepared in the at least solvent such as purified water and Dimethyl Sulphoxide (DMSO) at stock concentration of 0.05 %, at a speed in a range of 800-1200 rpm for 10-15 min at 35-38 °C.
2. The active ingredients at their working concentration i.e. 0.003125 % of TA and 0.005 % of HA were mixed together along with preservatives such as methyl and propyl paraben at 400-800 rpm for 10-15 min at 35-40 °C.

3. Then, the required quantity of water phase components such as sandalwood distillate water and colloidal oatmeal were weighed in a heat proof glass.
4. In another set, oil phase components such as castor oil, bees wax, xanthum gum and glyceryl stearate were weighed separately.
5. Both sets were incubated in water bath at 70 °C for 45 min, separately.
6. After incubation, at least one water phase component mixed with at least one oil phase component at 900-1100 rpm for 5-10 minutes at a temperature range of 40-45 °C to obtain a first mixture of water and oil phase.
7. The active ingredients were blended with the first mixture at 1000 rpm for 2 - 10 minutes at a temperature range of 40-45 °C, to obtain the composition.
8. The formulated water in oil base lotion was stored at room temperature, to obtain the formulation.
Advantages of the present disclosure
[097] The present disclosure discloses a composition comprising tetradecanoic acid (TA) and hexadecanoic acid (HA), wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006% with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01% with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1. The combination of the phytochemicals provides a synergistic effect against the biofilm formation in the species of Candida genus such as C. albicans and C. tropicalis, even by using these phytochemicals at a very low concentration. The efficacy of the individual phytochemical is increased by the combinatorial strategy of using the combination of the composition as disclosed in the present disclosure. This combinatorial strategy of using the combination of phytochemicals helps in overcoming the toxic side effects and hence do not affect the viability of cells.

Additionally, this combinatorial strategy of using the combination of phytochemicals helps in reducing the probability of developing multidrug resistance. The composition of the present disclosure is used in preparing the formulation. The formulation can be further used commercially in the form of topical cream and topical lotion. Overall the present disclosure provides a cost-effect, efficient, and non-toxic composition,

A composition comprising:
a. tetradecanoic acid; and
b. hexadecanoic acid,
wherein tetradecanoic acid is having a weight percentage in a range of 0.001-0.006% with respect to the composition, hexadecanoic acid is having a weight percentage in a range of 0.002-0.01% with respect to the composition and tetradecanoic acid to hexadecanoic acid has a weight ratio in a range of 0.350:1 to 1:1. The composition as claimed in claim 1, wherein the composition further comprises at least one water phase component.
The composition as claimed in claim 1, wherein the composition further comprises at least one oil phase component.
The composition as claimed in claim 1, wherein the composition further comprises a combination of at least one water phase and at least one oil phase component.
The composition as claimed in claim 2, wherein the at least one water phase component is selected from a group consisting of hydrosols, humectants, and combinations thereof.
The composition as claimed in claim 3, wherein the at least one oil phase component is selected from a group consisting of preservatives, emollients, structurants, thickeners, emulsifiers, and combinations thereof. The composition as claimed in claim 5, wherein the hydrosols have a weight percentage in a range of 50-80% with respect to the composition, humectants have a weight percentage in a range of 1-10% with respect to the composition. The composition as claimed in claim 6, wherein the preservatives have a weight percentage in range of 0.001-0.5%) with respect to the composition, emollients have a weight percentage in a range of 0.05-20%) with respect to the

composition, structurants have a weight percentage in a range of 0.01-7% with respect to the composition, thickeners have a weight percentage in a range of 0.05 to 5% with respect to the composition and emulsifiers have a weight percentage in a range of 1 to 10% with respect to the composition. A process for preparing the composition as claimed in claim 1, said process comprising: contacting tetradecanoic acid with hexadecanoic acid in presence of at least one solvent, at a speed in a range of 800-1200 rpm at a temperature in a range of 32°C to 40°C for a time period in a range of 5 to 20 minutes to obtain the composition. 3. A process for preparing the composition as claimed in any one of the claims 2-4, said process comprising:
a. contacting tetradecanoic acid with hexadecanoic acid in presence of an
oil phase component at a speed in a range of 400-800 rpm for a time in
a range of 10-15 minutes at a temperature in a range of 35-40°C to obtain
a first mixture;
b. contacting at least one water phase component with at least one oil
phase component at a speed in a range of 900-1100 rpm for a time period
in a range of 5-10 minutes at a temperature in a range of 40-45°C, to
obtain a second mixture; and
c. contacting the first mixture and the second mixture at a speed in a range
of 900-1100 rpm for a time period in a range of 2-10 minutes at a
temperature in a range of 40-45°C to obtain the composition.
1. The process as claimed in any one of the claims 9 to 10, wherein the process further comprises at least one solvent, wherein the at least one solvent is selected from a group consisting of water, dimethyl sulphoxide, isopropanol, chloroform, hexane, carbon disulphide and combinations thereof.

The composition as claimed in anyone of the claims 1 to 8, wherein the composition exhibits inhibitory activity against biofilm formation in species of Candida genus.